Fingerprint quality control method of jin song zhiyang composition

Abstract

The application discloses a kind of Jin Song Zhiniao composition fingerprint quality control method, provide five characteristic fingerprints of Jin Song Zhiniao composition, especially its lotion quality consistency evaluation method, using macro quantitative similarity to control the range of Jin Song Zhiniao lotion preparation total drug efficacy material and quality consistency, establish the five characteristic fingerprint control mode of Jin Song Zhiniao lotion standard preparation. Among them, gradient elution is carried out by high performance liquid chromatograph and is determined;High performance liquid chromatography conditions are as follows: chromatographic column: C18 chromatographic column;Detection wavelength: 220 nm, 234 nm, 250 nm, 280 nm, 344 nm;Mobile phase: mobile phase A is 0.2 % phosphoric acid aqueous solution containing 0.005 mol / L sodium heptanesulfonate, mobile phase B is acetonitrile-methanol solution with volume ratio of 9:1, gradient elution is adopted.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of quality control of traditional Chinese medicine, and particularly relates to a fingerprint spectrum quality control method of a Jinsong Zhiyang composition. BACKGROUND

[0002] Jinsong Zhiyang lotion (Guojiazhunzi B20020210) is composed of Baixianpi, Qianliuang and other 9 kinds of traditional Chinese medicines, has the effects of clearing heat and removing dampness, killing insects and relieving itching, and is used for treating adult female vulvitis, damp-heat pruritus, and external genital and skin itching. Fingerprint spectrum technology is the most effective means for controlling the quality of complex systems globally, and has important significance in the determination of components and authenticity identification of traditional Chinese medicine or natural medicine quality. People have gradually realized that the quality control method formed only for a certain chemical component cannot properly reflect the internal quality of the product, and the effective means to solve this problem is to establish an innovative quality control method and a characteristic fingerprint spectrum control mode of standard preparation, and to strengthen the scientificity and standardization of traditional Chinese medicine quality evaluation. Therefore, it is particularly important to construct a multi-dimensional fingerprint spectrum (high performance liquid chromatography fingerprint spectrum, UV fingerprint spectrum, Fourier infrared fingerprint spectrum, electrochemical fingerprint spectrum and DSC fingerprint spectrum) of Jinsong Zhiyang lotion and to use five standard fingerprint spectrum evaluation modes of standard preparation to perform multi-dimensional quality control on Jinsong Zhiyang lotion.

[0003] Because traditional Chinese medicine has complexity, the inventor overcomes the technical problems, and develops five characteristic fingerprint spectra and quality consistency evaluation methods of Jinsong Zhiyang lotion standard preparation. The methods are overall quality control based on all components such as components with large polarity, medium polarity and low polarity. SUMMARY

[0004] The purpose of the present application is to provide a fingerprint spectrum quality control method of Jinsong Zhiyang composition. The present application provides five characteristic fingerprint spectra and quality consistency evaluation methods of Jinsong Zhiyang lotion standard preparation, and uses macro-quantitative similarity to control the range of total medicinal material of Jinsong Zhiyang lotion preparation and the quality consistency.

[0005] The technical content of the present application is as follows:

[0006] The fingerprint spectrum quality control method of the Jinsong Zhiyang composition comprises gradient elution and determination by a high performance liquid chromatograph;

[0007] The high performance liquid chromatography conditions are as follows:

[0008] The chromatographic column is a C18 chromatographic column;

[0009] The detection wavelength is 220 nm, 234 nm, 250 nm, 280 nm and 344 nm;

[0010] The mobile phase A is 0.2% phosphoric acid aqueous solution containing 0.005 mol / L sodium heptane sulfonate, and the mobile phase B is acetonitrile-methanol solution with a volume ratio of 9:1, and gradient elution is adopted, and the elution program is as follows:

[0011]

[0012]

[0013] The C18 chromatographic column is a COSMOSIL 5C18-MS-II packed column with a specification of 5 μm and 250 mm x 4.6 mm, the column temperature is 30-40 ℃, and the flow rate of the mobile phase is 0.8-1.5 mL / min.

[0014] The composition is made of the following medicinal materials: Senecio scandens Buchen, Dictamnus dasycarpus Turcz, Eucalyptus robusta Smith, Lonicera japonica, Scutellaria baicalensis Georgi, Smilax glabra, Cnidium monnieri, Pinus massoniana, and Glycyrrhiza uralensis.

[0015] When the composition is prepared into the Pinus massoniana lotion for relieving itching, the preparation method of the test sample solution comprises the following steps:

[0016] The Pinus massoniana lotion for relieving itching is directly filtered through a 0.45 μm filter membrane, and the filtered solution is taken to obtain the test sample solution.

[0017] The preparation method of the reference sample solution comprises the following steps:

[0018] The reference sample solution is prepared by accurately weighing a certain amount of gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonoside, fraxidin, and cnidimmonnieri, and adding methanol to prepare a mixed reference sample solution containing 100-150 μg of gallic acid, 40-60 μg of chlorogenic acid, 4-6 μg of isochlorogenic acid C, 200-250 μg of baicalin, 20-40 μg of wogonoside, 50-90 μg of fraxidin, and 30-50 μg of cnidimmonnieri per 1 ml, and shaking well.

[0019] The process of establishing the fingerprint spectrum is as follows:

[0020] 10 μL of the reference sample solution and the test sample solution are injected into the liquid chromatograph for determination, and the chromatogram is recorded to determine the gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonoside, fraxidin, and cnidimmonnieri in the test sample chromatogram, with baicalin as the reference peak; detection is performed at 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm to obtain 17, 30, 17, 16, and 10 characteristic peaks, respectively, the macro qualitative similarity of the characteristic spectrum of the test sample and the standard characteristic spectrum of the reference sample solution is not less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0021] The characteristic peaks of the standard preparation fingerprint spectrum detected at 220 nm are as follows:

[0022]

[0023]

[0024] The characteristic peaks of the standard preparation fingerprint spectrum detected at 234 nm are as follows:

[0025]

[0026] The characteristic peaks of the standard preparation fingerprint spectrum detected at 250 nm are as follows:

[0027]

[0028]

[0029] The characteristic peaks of the standard preparation fingerprint spectrum detected at 280 nm are as follows:

[0030]

[0031] The characteristic peaks of the standard preparation fingerprint spectrum detected at 344 nm are as follows:

[0032]

[0033] The fingerprint spectrum quality control method of the Jinsong Zhiyang composition further comprises a UV detection method for the Jinsong Zhiyang lotion, comprising the following steps:

[0034] a. Preparation of test sample solution

[0035] Dilute the Jinsong Zhiyang lotion sample by 10 times, pass through a 0.45 μm filter membrane, and take the filtrate to obtain the test sample solution;

[0036] b. UV chromatographic conditions

[0037] Replace the chromatographic column with a hollow polyether ether ketone tube 5000 mm x 0.12 mm; the mobile phase A is a 0.2% phosphoric acid aqueous solution containing 0.005 mol / L sodium heptanesulfonate, the mobile phase B is a volume ratio of 9:1 acetonitrile-methanol solution, the mobile phase A and B are gradient eluted at a ratio of 1:1; the detection wavelength is 220 nm, 234 nm, 250 nm, 280 nm, 344 nm, the column temperature is 40-50℃, and the flow rate is 0.4-0.6 mL per minute;

[0038] c. UV fingerprint spectrum inspection method

[0039] Take 2 μL of the reference solution and the test sample solution, inject into the liquid chromatograph, measure and record the chromatogram and spectrum, the macro qualitative similarity of the test sample UV characteristic spectrum and the standard characteristic spectrum of the UV control sample solution should not be less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0040] The fingerprint spectrum quality control method of the Jinsong Zhiyang composition further comprises an FT-IR fingerprint spectrum detection method for the Jinsong Zhiyang lotion, comprising the following steps:

[0041] a. Preparation of test sample

[0042] The Jinsong Zhiyang lotion is dried into powder, the powder is dried in a blast drying oven at 100°C for 30 min, taken out and stored in a glass desiccator containing phosphorus pentoxide desiccant; the sample is placed in a marble mortar at a weight ratio of 1 / 12 with KBr, ground in the same direction, mixed uniformly, then the mixture is compressed under a pressure of 15 MPa for 3 min to form a uniform transparent wafer, which is taken out, and the FT-IR characteristic spectrum of the Jinsong Zhiyang lotion is obtained.

[0043] b. Preparation of blank sample

[0044] Without adding the Jinsong Zhiyang lotion sample, only KBr is used, and the remaining steps are prepared in parallel, compressed under a pressure of 15 MPa for 3 min to form a uniform thin transparent potassium bromide wafer, which is taken out, and the FT-IR characteristic spectrum of the Jinsong Zhiyang lotion is obtained.

[0045] c. Experimental conditions

[0046] The Jinsong Zhiyang lotion test sample wafer and the blank KBr wafer are respectively placed into an iCAN9 Fourier transform infrared spectrometer for measurement, and the infrared absorption spectrum of the mixture in the range of 4000-400 cm -1 The macro qualitative similarity of the test sample FT-IR characteristic spectrum and the standard characteristic spectrum of the FR-IR control sample solution should not be less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0047] The fingerprint spectrum quality control method of the Jinsong Zhiyang composition further comprises an electrochemical fingerprint spectrum detection method for the Jinsong Zhiyang lotion, comprising the following steps:

[0048] a. Preparation of chemical oscillation system solution:

[0049] Prepare 3 mol / L sulfuric acid solution, 0.2 mol / L sulfuric acid solution, 0.4 mol / L malonic acid solution, 0.02 mol / L cerium ammonium sulfate solution, and 0.3 mol / L potassium bromate solution for standby;

[0050] b. Preparation of test sample

[0051] Precise measuring 1.0 mL of Jinsong Zhiyang lotion sample into a 10 mL volumetric flask, adding purified water to constant volume, shaking well, and mixing thoroughly to obtain a test sample;

[0052] c. Experimental conditions

[0053] Hg / Hg2Cl2 / KCl as a reference electrode, platinum electrode as a working electrode, the speed of DF-101S heat collection type constant temperature heating magnetic stirrer is 400 revolutions per minute, the water bath temperature is 37 DEG C, and the addition amount of each solution in the chemical oscillation system is as follows:

[0054] Solution 3 mol / L sulfuric acid 0.4 mol / L malonic acid 0.02 mol / L cerium sulfate ammonium 0.3 mol / L potassium bromate Sample / mL Added (mL) 12.0 6.0 3.0 3.0 1.5

[0055] d. Electrochemical fingerprint examination method

[0056] 1.5 mL of Jinsong Zhiyang lotion test sample solution is taken to determine the electrochemical fingerprint, and the fingerprint is recorded, the macro qualitative similarity of the electrochemical characteristic fingerprint of the test sample and the electrochemical standard characteristic fingerprint should not be less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0057] The fingerprint quality control method of the Jinsong Zhiyang composition also includes a DSC fingerprint detection method for Jinsong Zhiyang lotion, comprising the following steps:

[0058] a. Detection conditions

[0059] A gradient temperature program is used: 40-500 DEG C is 15 DEG C / min, and 500 DEG C is constant temperature for 5 min. The atmosphere is air;

[0060] b. DSC fingerprint examination method

[0061] Jinsong Zhiyang lotion is dried into powder, 6 mg of Jinsong Zhiyang lotion dry powder is accurately weighed in an aluminum crucible, and is placed into a thermal balance for measurement, and the heat flow rate change is recorded, the macro qualitative similarity of the DSC characteristic spectrum of the test sample and the standard characteristic spectrum of the DSC control sample solution should not be less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0062] The beneficial effects of the present application are:

[0063] The present application realizes the Jinsong Zhiyang composition, especially the whole quality control mode and control method of the lotion preparation thereof, and establishes five characteristic fingerprint control modes of the standard preparation of Jinsong Zhiyang lotion. The average of the macro qualitative similarity of the five fingerprints should be greater than 0.90, and the average of the macro quantitative similarity of the five fingerprints should be between 80% and 120%.

[0064] The present application establishes the quality standard of non-volatile substances based on multi-index components, can accurately control and evaluate the quality of Jin Song Zhi Yang lotion, and makes up for the deficiency of the current quality control method.

[0065] The method of the present application has good feasibility, stability and reproducibility. BRIEF DESCRIPTION OF DRAWINGS

[0066] Figure 1 HPLC characteristic fingerprint of Jin Song Zhi Yang lotion standard preparation is obtained by using 220nm detection;

[0067] Figure 2 HPLC characteristic fingerprint of Jin Song Zhi Yang lotion standard preparation is obtained by using 234nm detection;

[0068] Figure 3 HPLC characteristic fingerprint of Jin Song Zhi Yang lotion standard preparation is obtained by using 250nm detection;

[0069] Figure 4 HPLC characteristic fingerprint of Jin Song Zhi Yang lotion standard preparation is obtained by using 280nm detection;

[0070] Figure 5 HPLC characteristic fingerprint of Jin Song Zhi Yang lotion standard preparation is obtained by using 344nm detection;

[0071] Figure 6 UV standard characteristic quantum fingerprint of Jin Song Zhi Yang lotion standard preparation (33 quantum fingerprint peaks are obtained by combining 5 points);

[0072] Figure 7 FT-IR standard characteristic quantum fingerprint of Jin Song Zhi Yang lotion standard preparation (76 quantum fingerprint peaks are obtained by combining 5 points);

[0073] Figure 8 Electrochemical standard characteristic fingerprint of Jin Song Zhi Yang lotion standard preparation is obtained by using electrochemical workstation detection;

[0074] Figure 9 DSC standard characteristic quantum fingerprint of Jin Song Zhi Yang lotion standard preparation is obtained by using DSC detection;

[0075] Figure 10 DSC standard characteristic quantum fingerprint of Jin Song Zhi Yang lotion (137 quantum fingerprint peaks are obtained by combining 50 points). DETAILED DESCRIPTION

[0076] The present application will be further described below by examples. It should be understood that the examples of the present application are used to illustrate the present application rather than limit the present application.

[0077] Example 1

[0078] The composition is made of Senecio scandens Buchen, Dictamnus dasycarpus Turcz, Eucalyptus tereticornis Smith, Lonicera japonica, Scutellaria baicalensis Georgi, Smilax glabra, Cnidium monnieri, Pinus massoniana, and Glycyrrhiza uralensis.

[0079] When the composition is prepared into the Jinsong Zhiyang lotion, the preparation method of the test sample solution comprises the following steps.

[0080] The Jinsong Zhiyang lotion is directly filtered through a 0.45 μm filter membrane, and the filtered solution is taken to obtain the test sample solution.

[0081] The preparation method of the control sample solution comprises the following steps.

[0082] The appropriate amounts of gallic acid control sample, chlorogenic acid control sample, isochlorogenic acid C control sample, baicalin control sample, wogonoside control sample, fraxinellone control sample, and cnidimide control sample are precisely weighed and determined, and methanol is added to prepare a mixed control sample solution containing 100 μg of gallic acid, 40 μg of chlorogenic acid, 4 μg of isochlorogenic acid C, 200 μg of baicalin, 20 μg of wogonoside, 50 μg of fraxinellone, and 30 μg of cnidimide per 1 ml, and the mixture is shaken to obtain the mixed control sample solution.

[0083] The fingerprint spectrum is established in the following process.

[0084] The fingerprint spectrum quality control method of the Jinsong Zhiyang composition comprises gradient elution and determination by a high-performance liquid chromatograph.

[0085] The high-performance liquid chromatography conditions are as follows.

[0086] The chromatographic column is a COSMOSIL 5C18-MS-II packed column with a specification of 5 μm and 250 mm×4.6 mm, the column temperature is 30℃, and the flow rate of the mobile phase is 0.8 mL / min.

[0087] The detection wavelength is 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm.

[0088] The mobile phase A is 0.2% phosphoric acid aqueous solution containing 0.005 mol / L sodium heptanesulfonate, the mobile phase B is a solution of acetonitrile-methanol with a volume ratio of 9:1, gradient elution is adopted, and the elution program is as follows.

[0089]

[0090] Take 10 μL of the reference substance solution and 10 μL of the test sample solution, inject into the liquid chromatograph, determine, record the chromatogram, determine the gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonoside, fraxidin, and cnidilide in the chromatogram of the test sample, take baicalin as the reference substance peak; detect at 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm, and obtain 17, 30, 17, 16, and 10 characteristic peaks respectively, the macro qualitative similarity of the characteristic spectrum of the test sample to the standard characteristic spectrum of the control substance solution is not less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0091] Example 2

[0092] The composition is made of Senecio scandens Buchen, Dictamnus dasycarpus Turcz, Eucalyptus robusta Smith, Lonicera japonica, Scutellaria baicalensis Georgi, Smilax glabra, Cnidium monnieri, Pinus massoniana, and Glycyrrhiza uralensis.

[0093] When the composition is prepared into the Jin Song Zhi Yang lotion, the preparation method of the test sample solution comprises the following steps.

[0094] The Jin Song Zhi Yang lotion is directly filtered through a 0.45 μm filter membrane, and the filtered solution is taken to obtain the test sample solution.

[0095] The preparation method of the control substance solution comprises the following steps.

[0096] Take appropriate amounts of gallic acid control substance, chlorogenic acid control substance, isochlorogenic acid C control substance, baicalin control substance, wogonoside control substance, fraxidin control substance, and cnidilide control substance respectively, accurately weigh and determine, add methanol to prepare a mixed control substance solution containing 150 μg of gallic acid, 60 μg of chlorogenic acid, 6 μg of isochlorogenic acid C, 250 μg of baicalin, 40 μg of wogonoside, 50 μg of fraxidin, and 30 μg of cnidilide per 1 ml, and shake well.

[0097] The process of establishing the fingerprint spectrum is as follows.

[0098] The fingerprint spectrum quality control method of the Jin Song Zhi Yang composition comprises gradient elution and determination by a high performance liquid chromatograph.

[0099] The high performance liquid chromatography conditions are as follows.

[0100] The chromatographic column is a COSMOSIL 5C18-MS-II packed column with a specification of 5 μm and 250 mm x 4.6 mm, the column temperature is 40℃, and the flow rate of the mobile phase is 1.5 mL / min.

[0101] The detection wavelengths are 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm.

[0102] The mobile phase A is 0.2% phosphoric acid aqueous solution containing 0.005 mol / L sodium heptane sulfonate, and the mobile phase B is acetonitrile-methanol solution with a volume ratio of 9:1, gradient elution is adopted, and the elution program is as follows:

[0103]

[0104] The reference solution and the test solution are each 10 μL, injected into the liquid chromatograph, measured, and the chromatogram is recorded to determine the gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonoside, fraxidin, and cnidilide in the test product chromatogram, with baicalin as the reference peak; detection is performed at 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm, and 17, 30, 17, 16, and 10 characteristic peaks are obtained respectively, the macro qualitative similarity of the test product characteristic spectrum and the standard characteristic spectrum of the control solution should not be less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0105] Example 3

[0106] The composition is made of the following medicinal materials: groundsel, barks of Chinese globeandleaf, eucalyptus tereticornis smith, honeysuckle, scutellaria baicalensis, smilax glabra, cnidium monnieri, pine needles, and licorice.

[0107] When the composition is prepared into the Jin Song Zhi Ya lotion, the preparation method of the test product solution comprises the following steps:

[0108] The Jin Song Zhi Ya lotion is directly filtered through a 0.45 μm filter membrane, and the filtered solution is taken as the test product solution.

[0109] The preparation method of the control solution comprises the following steps:

[0110] The gallic acid control, chlorogenic acid control, isochlorogenic acid C control, baicalin control, wogonoside control, fraxidin control, and cnidilide control are respectively accurately weighed and determined, and methanol is added to prepare a mixed control solution containing 120 μg of gallic acid, 50 μg of chlorogenic acid, 5 μg of isochlorogenic acid C, 220 μg of baicalin, 30 μg of wogonoside, 70 μg of fraxidin, and 40 μg of cnidilide per 1 ml, and then shaken to obtain the mixed control solution.

[0111] The process of establishing the fingerprint spectrum is as follows:

[0112] The fingerprint spectrum quality control method of the Jin Song Zhi Ya composition comprises gradient elution and measurement by a high-performance liquid chromatograph;

[0113] The high-performance liquid chromatography conditions are as follows:

[0114] Chromatographic column: COSMOSIL 5C18-MS-II packed column, specifications: 5 μm, 250 mm x 4.6 mm; the column temperature was 35℃; the flow rate of the mobile phase was 1 mL / min;

[0115] Detection wavelength: 220 nm, 234 nm, 250 nm, 280 nm, 344 nm;

[0116] Mobile phase: mobile phase A was 0.2% phosphoric acid aqueous solution containing 0.005 mol / L sodium heptanesulfonate, mobile phase B was acetonitrile-methanol solution with a volume ratio of 9:1, gradient elution was used, and the elution program was as follows:

[0117]

[0118] Each 10 μL of the reference substance solution and the test sample solution was injected into the liquid chromatograph for determination, the chromatogram was recorded, and the gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonoside, jujubinone, and cnidilide in the test sample chromatogram were determined with baicalin as the reference substance peak; 17, 30, 17, 16, and 10 characteristic peaks were obtained at 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm, respectively; the macro qualitative similarity of the test sample characteristic spectrum and the standard characteristic spectrum of the control substance solution was not less than 0.90, and the macro quantitative similarity should be between 80% and 120%.

[0119] Example 4

[0120] The present example provides a high-performance liquid chromatography characteristic fingerprint and quality consistency evaluation method for the standard preparation of Jinsong Zhiyang lotion, which comprises the following steps:

[0121] S1 Preparation of test sample solution

[0122] An appropriate amount of Jinsong Zhiyang lotion sample was directly filtered through a 0.45 μm filter membrane, and the filtrate was collected to obtain the test sample solution;

[0123] S2 Preparation of control substance solution

[0124] An appropriate amount of gallic acid control substance, chlorogenic acid control substance, isochlorogenic acid C control substance, baicalin control substance, wogonoside control substance, jujubinone control substance, and cnidilide control substance was precisely weighed and added to methanol to prepare a mixed control substance solution containing 123.8 μg of gallic acid, 45.5 μg of chlorogenic acid, 4.4 μg of isochlorogenic acid C, 226 μg of baicalin, 28.1 μg of wogonoside, 73.2 μg of jujubinone, and 39.7 μg of cnidilide per 1 ml, and then the mixture was shaken to obtain the control substance solution;

[0125] S3 HPLC chromatographic conditions

[0126] Column: octadecylsilane-bonded silica gel as the filler (COSMOSIL 5C18-MS-II, column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); mobile phase A: 0.2% phosphoric acid-water solution (containing 0.005 mol / L sodium heptanesulfonate), mobile phase B: acetonitrile-methanol (9:1) solution, gradient elution according to the table below; detection wavelength: 220 nm, 234 nm, 250 nm, 280 nm, 344 nm; column temperature: 35 °C, flow rate: 1.0 mL per minute, injection volume: 10 μL;

[0127] Table 1 Gradient elution procedure table for HPLC fingerprint of Jinsong Zhiyang lotion

[0128]

[0129] S4 HPLC fingerprint inspection method

[0130] Inject 10 μL of the reference solution and 10 μL of the test solution into the liquid chromatograph, determine and record the chromatogram. Determine the peaks of gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonoside, jujubinone, and cnidilide in the chromatogram of the test sample, taking baicalin as the reference peak. The chromatogram of the test sample should present thirty characteristic peaks corresponding to the standard characteristic chromatogram of the control sample solution. The peaks corresponding to the reference are S peaks. Evaluate the test sample characteristic chromatogram and the standard characteristic chromatogram of the control sample solution using the

Traditional Chinese Medicine Quantitative Fingerprint Software

[0131] S5 HPLC index component quantitative method

[0132] Use the standard curve method to quantitatively determine the seven compound components in Jinsong Zhiyang lotion. The linearity, range, detection limit, and quantitative limit of the seven compound components in Jinsong Zhiyang lotion are shown in Table 2.

[0133] Table 2 Linearity, range, detection limit, and quantitative limit of the seven index compounds

[0134]

[0135] Further, the detection wavelengths in step S5 include but are not limited to 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm, which include any wavelength within 190-400 nm collected by DAD for fingerprint detection;

[0136] Further, the number of common fingerprint peaks in step S5 includes but is not limited to 17, 30, 17, 16, and 10. Fingerprint peaks with a signal 5 times or more than the noise can be considered as effective fingerprint peaks;

[0137] As Figure 1 shown, the standard preparation fingerprint data of this example detected at 220 nm is as follows:

[0138]

[0139] As Figure 2 shown, the standard preparation fingerprint data of this example detected at 234 nm is as follows:

[0140]

[0141]

[0142] As Figure 3 shown, the standard preparation fingerprint data of this example detected at 250 nm is as follows:

[0143]

[0144] As Figure 4 shown, the standard preparation fingerprint data of this example detected at 280 nm is as follows:

[0145]

[0146] As Figure 5 shown, the standard preparation fingerprint data of this example detected at 344 nm is as follows:

[0147]

[0148] Example 5

[0149] The present example provides a UV fingerprint and quality consistency evaluation method for Jin Song Zhi Yang lotion standard preparation, comprising the following steps:

[0150] US1 preparation of test sample solution

[0151] Dilute Jin Song Zhi Yang lotion sample 10 times, pass through 0.45 μm filter membrane, take the filtrate to obtain the test sample solution;

[0152] US2 UV chromatographic conditions

[0153] The hollow polyether ether ketone (PEEK) tube (5000 mm x 0.12 mm) is used instead of the chromatographic column; 0.2% phosphoric acid-water solution (containing 0.005 mol / L sodium heptane sulfonate) is used as the mobile phase A, acetonitrile-methanol (9:1) solution as the mobile phase B, gradient elution is carried out with the mobile phase ratio of 1:1; the detection wavelength is 220 nm, 234 nm, 250 nm, 280 nm, 344 nm, the column temperature is 35°C, the flow rate is 0.5 mL per minute, and the injection amount is 2 μL. The absorption spectrum of the detected sample is recorded at the wavelength of 190 nm to 400 nm.

[0154] US3 UV fingerprint inspection method

[0155] 2 μL of the test sample solution is precisely taken and injected into the liquid chromatograph for determination, and the chromatogram and spectrum are recorded, as shown in Figure 6 The UV characteristic spectrum (quantum fingerprint spectrum) of the test sample and the standard characteristic spectrum (quantum fingerprint spectrum) of the UV control sample solution are evaluated by using the

Traditional Chinese Medicine Quantitative Fingerprint Software

[0156] Example 6

[0157] The present embodiment provides a FT-IR characteristic fingerprint spectrum and quality consistency evaluation method of Jin Song Zhi Yang lotion standard preparation, which comprises the following steps:

[0158] FS1 preparation of test sample

[0159] The Jin Song Zhi Yang lotion is dried into powder, and the powder is dried in a blast drying oven at 100°C for 30 min. The sample is taken out and placed in a glass desiccator containing phosphorus pentoxide desiccant to minimize water interference and obtain high-quality spectrum (higher intensity and lower noise interference). The sample is placed in a marcasite mortar at a ratio of 1 / 12 (w / w) with KBr, ground in the same direction, and mixed uniformly. Then the mixture (0.121 g) is compressed under a pressure of 15 MPa for 3 min to form a uniform transparent wafer, which is taken out to obtain the sample.

[0160] FS2 preparation of blank sample

[0161] Without adding the Jin Song Zhi Yang lotion sample, the remaining steps are prepared in parallel. A uniform thin transparent potassium bromide wafer is formed under a pressure of 15 MPa for 3 min, which is taken out to obtain the sample.

[0162] FS3 experimental conditions

[0163] The test sample wafer of Jin Song Zhi Yang lotion and the blank potassium bromide wafer were respectively put into iCAN9 Fourier transform infrared spectrometer for determination, and the infrared absorption spectrum of the mixture in the range of 4000-400 cm-1 wave number was recorded, see Figure 7 The sample data was recorded and exported in *.CSV file format, and evaluated by

Traditional Chinese Medicine Quantitative Fingerprint Software

[0164] Example 7

[0165] The present embodiment provides an electrochemical characteristic fingerprint spectrum and quality consistency evaluation method of Jin Song Zhi Yang lotion standard preparation, which comprises the following steps:

[0166] Preparation of ES1 chemical oscillation system solution

[0167] Preparation of 3 mol / L sulfuric acid solution: 162 mL of 98% concentrated sulfuric acid and 834 mL of purified water were respectively measured, and the purified water was placed in a beaker. Then the measured 98% concentrated sulfuric acid was slowly added to the water along the cup wall while stirring. After the addition was completed, it was cooled to room temperature.

[0168] Preparation of 0.2 mol / L sulfuric acid solution: according to the subsequent required amount, 0.2 mol / L sulfuric acid can be prepared by diluting 15 times of the prepared 3 mol / L sulfuric acid.

[0169] Preparation of 0.4 mol / L malonic acid solution: 10.4060 g of malonic acid was accurately weighed and placed in a 250 mL volumetric flask. Then 0.2 mol / L sulfuric acid was added to constant volume, and shaken well.

[0170] Preparation of 0.02 mol / L cerium ammonium sulfate solution: 3.3428 g of cerium ammonium sulfate was accurately weighed and placed in a 250 mL volumetric flask. Then 0.2 mol / L sulfuric acid was added to constant volume, and shaken well.

[0171] Preparation of 0.3 mol / L potassium bromate solution: 12.5250 g of potassium bromate was accurately weighed and placed in a 250 mL volumetric flask. Then purified water was added to constant volume, and shaken well.

[0172] Preparation of ES2 test sample

[0173] 1.0 mL of Jin Song Zhi Yang lotion sample was accurately measured and placed in a 10 mL volumetric flask. Then purified water was added to constant volume, and shaken well to mix thoroughly, to prepare the test sample solution.

[0174] ES3 experimental conditions

[0175] The Hg / Hg2Cl2 / KCl is used as reference electrode, the platinum electrode is used as working electrode, the rotating speed of DF-101S heat collection type constant temperature heating magnetic stirrer is 400 r / min, the water bath temperature is 37℃, and the adding amount of each solution in the chemical oscillation system is shown in the following table.

[0176] Solution 3 mol / L sulfuric acid 0.4 mol / L malonic acid 0.02 mol / L cerium sulfate ammonium 0.3 mol / L potassium bromate Sample / mL Added (mL) 12.0 6.0 3.0 3.0 1.5

[0177] ES4 fingerprint inspection method

[0178] 1.5 mL of the test solution of Jinsong Zhiyu lotion is taken, the electrochemical fingerprint is determined, seen in Figure 8 , and the fingerprint is recorded. After the *.CSV format file is exported by using

CHI660E Electrochemical Workstation

Traditional Chinese Medicine Quantitative Fingerprint Software

Traditional Chinese Medicine Quantum Software

[0179] Example 8

[0180] The embodiment provides a DSC characteristic fingerprint and quality consistency evaluation method of a Jinsong Zhiyu lotion standard preparation, which comprises the following steps:

[0181] DS1 detection condition

[0182] A gradient temperature rising program is used: 40-500℃ is 15℃ / min, and the temperature is kept at 500℃ for 5 min. The atmosphere is air;

[0183] DS2 DSC fingerprint determination method

[0184] The Jinsong Zhiyu lotion is dried into powder. 6 mg of sample powder is accurately weighed in an aluminum crucible, is placed into a thermal balance for determination, and the heat flow rate change is recorded, seen in Figure 9 and 10 , and the sample data is recorded and exported in *.CSV file format, and is evaluated by using

Traditional Chinese Medicine Quantitative Fingerprint Software

[0185] The above is the embodiment of the present application. The above embodiment and the specific parameters in the embodiment are only for clearly describing the application verification process, and are not used to limit the patent protection scope of the present application. The patent protection scope of the present application is still subject to its claims, and any equivalent structural changes made by using the contents of the specification and drawings of the present application should also be included in the protection scope of the present application.

Claims

1. A method for quality control of the fingerprint spectrum of a Jinsong antipruritic composition, characterized in that: This includes gradient elution and determination using high-performance liquid chromatography; The high-performance liquid chromatography conditions are as follows: Chromatographic column: C18 column; the C18 column is a COSMOSIL 5C18-MS-Ⅱ packed column with dimensions of 5μm and 250mm × 4.6mm. Detection wavelengths: 220 nm, 234 nm, 250 nm, 280 nm, 344 nm; Mobile phase: Mobile phase A is an aqueous solution of 0.2% phosphoric acid containing 0.005 mol / L sodium heptanesulfonate; mobile phase B is an acetonitrile-methanol solution with a volume ratio of 9:

1. Gradient elution is used, and the elution program is as follows: ； The Jinsong antipruritic composition is made from the following medicinal materials: Senecio scandens, Dictamnus dasycarpus root bark, Eucalyptus macrocarpa leaves, Lonicera japonica, Scutellaria baicalensis, Smilax glabra, Cnidium monnieri, pine needles, and Glycyrrhiza uralensis. The preparation method of the test solution includes the following steps: take Jinsong antipruritic lotion and filter it directly through a 0.45 μm filter membrane, take the filtrate, and obtain the test solution; The method for preparing the reference solution includes the following steps: Accurately weigh gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonin, morrhubarb, and osthol, respectively, and prepare a mixed reference solution containing 100-150 μg gallic acid, 40-60 μg chlorogenic acid, 4-6 μg isochlorogenic acid C, 200-250 μg baicalin, 20-40 μg wogonin, 50-90 μg morrhubarb, and 30-50 μg osthol per ml in methanol. Shake well to obtain the final solution.

2. The fingerprint spectrum quality control method for the Jinsong antipruritic composition as described in claim 1, characterized in that: The flow rate of the mobile phase is 0.8-1.0 mL / min.

3. The fingerprint spectrum quality control method for the Jinsong antipruritic composition as described in claim 1, characterized in that: The fingerprint pattern establishment process is as follows: Take 10 μL each of the reference solution and the test solution, inject them into the liquid chromatograph, determine and record the chromatograms, and identify gallic acid, chlorogenic acid, isochlorogenic acid C, baicalin, wogonin, morrhurin, and osthol in the chromatogram of the test solution, using baicalin as the reference peak; detect at 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm, respectively, and obtain 17, 30, 17, 16, and 10 characteristic peaks, respectively. The macro-qualitative similarity between the characteristic chromatogram of the test solution and the standard characteristic chromatogram of the reference solution shall not be less than 0.90, and the macro-quantitative similarity shall be between 80% and 120%.

4. The fingerprint spectrum quality control method for the Jinsong antipruritic composition as described in claim 3, characterized in that: The characteristic peaks of the standard formulation fingerprint spectrum detected at 220 nm are as follows: ； The characteristic peaks of the standard formulation fingerprint spectrum detected at 234 nm are as follows: ； The characteristic peaks of the standard formulation fingerprint spectrum detected at 250 nm are as follows: ； The characteristic peaks of the standard formulation fingerprint spectrum detected at 280 nm are as follows: ； The characteristic peaks of the standard formulation fingerprint detected at 344 nm are as follows: 。 5. The fingerprint spectrum quality control method for the Jinsong antipruritic composition as described in claim 1, characterized in that: It also includes a UV detection method for Jinsong antipruritic lotion, comprising the following steps: a. Preparation of test solution Dilute the Jinsong antipruritic lotion sample 10 times, filter it through a 0.45 μm filter membrane, and collect the filtrate to obtain the test solution; b. UV chromatographic conditions A hollow polyetheretherketone (PEEK) tube (5000 mm × 0.12 mm) was used instead of a chromatographic column. Mobile phase A was an aqueous solution of 0.2% phosphoric acid containing 0.005 mol / L sodium heptanesulfonate, and mobile phase B was an acetonitrile-methanol solution with a volume ratio of 9:

1. Mobile phases A and B were eluted isocratically at a ratio of 1:

1. The detection wavelengths were 220 nm, 234 nm, 250 nm, 280 nm, and 344 nm. The column temperature was 40-50 ℃, and the flow rate was 0.4-0.6 mL per minute. c. UV fingerprinting method Take 2 μL each of the reference solution and the test solution, inject them into the liquid chromatograph, measure and record the chromatogram and spectrum. The macro-qualitative similarity between the UV characteristic spectrum of the test sample and the standard characteristic spectrum of the UV reference solution should not be less than 0.90, and the macro-quantitative similarity should be between 80% and 120%.

6. The fingerprint spectrum quality control method for the Jinsong antipruritic composition as described in claim 1, characterized in that: It also includes an FT-IR fingerprinting method for Jinsong antipruritic lotion, comprising the following steps: a. Preparation of test sample The Jin Song antipruritic lotion was dried into powder, and the powder was dried in a forced-air drying oven at 100℃ for 30 min. It was then removed and stored in a glass desiccator containing phosphorus pentoxide desiccant. The sample was placed in an agate mortar with KBr at a weight ratio of 1 / 12 and ground in the same direction until it was mixed evenly. The mixture was then compressed under a pressure of 15 MPa for 3 min to form a uniform transparent crystal. The crystal was then removed and the product was obtained. b. Prepare blank samples Without adding Jinsong antipruritic lotion sample, only KBr was used, and the remaining steps were prepared in parallel. The sample was pressed at 15 MPa for 3 min to form a uniform, thin, transparent potassium bromide crystal, which was then removed to obtain the product. c. Experimental conditions Take the test sample crystal of Jinsong Antipruritic Lotion and the blank KBr crystal separately and place them in an iCAN9 Fourier transform infrared spectrometer for measurement. Record the temperature of the mixture at 4000-400 cm⁻¹. -1 For the infrared absorption spectra in the wavenumber range, the macro-qualitative similarity between the FT-IR characteristic spectrum of the test sample and the standard characteristic spectrum of the FR-IR reference solution shall not be less than 0.90, and the macro-quantitative similarity shall be between 80% and 120%.

7. The fingerprint spectrum quality control method for the Jinsong antipruritic composition as described in claim 1, characterized in that: It also includes an electrochemical fingerprinting method for detecting Jinsong antipruritic lotion, comprising the following steps: a. Preparation of the chemical shaking system solution: Prepare 3 mol / L sulfuric acid solution, 0.2 mol / L sulfuric acid solution, 0.4 mol / L malonic acid solution, 0.02 mol / L cerium ammonium sulfate solution, and 0.3 mol / L potassium bromate solution for later use; b. Preparation of the test sample Accurately measure 1.0 mL of Jinsong antipruritic lotion sample and place it in a 10 mL volumetric flask. Add purified water to make up to the mark, shake well, and mix thoroughly to prepare the test sample. c. Experimental conditions Using Hg / Hg2Cl2 / KCl as the reference electrode and a platinum electrode as the working electrode, the DF-101S thermal collector-type constant temperature heating magnetic stirrer rotates at 400 rpm, and the water bath temperature is 37 ℃. The amounts of each solution added to the chemical oscillation system are shown in the table below. ； d. Electrochemical fingerprinting method Take 1.5 mL of the Jinsong Antipruritic Lotion test solution, determine the electrochemical fingerprint spectrum, and record the fingerprint spectrum. The macro-qualitative similarity between the electrochemical characteristic fingerprint spectrum of the test sample and the electrochemical standard characteristic fingerprint spectrum shall not be less than 0.90, and the macro-quantitative similarity shall be between 80% and 120%.

8. The fingerprint spectrum quality control method for the Jinsong antipruritic composition as described in claim 1, characterized in that: It also includes a DSC fingerprinting method for detecting Jinsong antipruritic lotion, comprising the following steps: a. Testing conditions A gradient heating program was used: 40-500 ℃ at a rate of 15 ℃ / min, followed by a constant temperature of 500 ℃ for 5 min, with air as the atmosphere; b. DSC fingerprint examination method Dry Jinsong antipruritic lotion into powder. Accurately weigh 6 mg of the dried powder into an aluminum crucible, place it in a thermobalance for determination, and record the change in heat flow rate. The macro-qualitative similarity between the DSC characteristic spectrum of the test sample and the standard characteristic spectrum of the DSC reference solution should not be less than 0.90, and the macro-quantitative similarity should be between 80% and 120%.

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